Designing conformational control of human tissue transglutaminase

Understanding the mechanisms of enzymes is crucial for our understanding of their role in biology and for designing methods to perturb or harness their activities for medical treatments, industrial processes, or biological engineering. One aspect of enzymes that makes them difficult to fully understand is that they are in constant motion, and these motions and the conformations adopted throughout these transitions often play a role in their function.

Traditionally, it has been difficult to isolate a protein in a particular conformation to determine what role each form plays in the reaction or biology of that enzyme. A new technology, computational protein design, makes the isolation of various conformations possible, and therefore is an extremely powerful tool in enabling a fuller understanding of the role a protein conformation plays in various biological processes.

One such protein that undergoes large structural shifts during different activities is human type II transglutaminase (TG2). TG2 is an enzyme that exists in two dramatically different conformational states: (1) an open, extended form, which is adopted upon the binding of calcium, and (2) a closed, compact form, which is adopted upon the binding of GTP or GDP. TG2 possess two separate active sites, each with a radically different activity. This open, calcium-bound form of TG2 is believed to act as a transglutaminse, where it catalyzes the formation of an isopeptide bond between the sidechain of a peptide-bound glutamine and a primary amine. The closed, GTP-bound conformation is believed to act as a GTPase. TG2 is also implicated in a variety of biological and pathological processes.

To better understand the effects of TG2’s conformations on its activities and pathological processes, we set out to design variants of TG2 isolated in either the closed or open conformations. We were able to design open-locked and closed-biased TG2 variants, and use these designs to unseat the current understanding of the activities and their concurrent conformations of TG2 and explore each conformation’s role in celiac disease models. This work also enabled us to help explain older confusing results in regards to this enzyme and its activities. The new model for TG2 activity has immense implications for our understanding of its functional capabilities in various environments, and for our ability to understand which conformations need to be inhibited in the design of new drugs for diseases in which TG2’s activities are believed to elicit pathological effects.

On innate and specific aspects of human language

Mixel Aurnague, Kepa Korta and Jesus M. Larrazabal (eds.)

Studies in nuclear reactor dynamics : I. The accuracy of point kinetics. II. The effect of delayed neutrons on the spectrum of the group-diffusion operator

This thesis is a theoretical work on the space-time dynamic behavior of a nuclear reactor without feedback. Diffusion theory with G-energy groups is used.

In the first part the accuracy of the point kinetics (lumped-parameter description) model is examined. The fundamental approximation of this model is the splitting of the neutron density into a product of a known function of space and an unknown function of time; then the properties of the system can be averaged in space through the use of appropriate weighting functions; as a result a set of ordinary differential equations is obtained for the description of time behavior. It is clear that changes of the shape of the neutron-density distribution due to space-dependent perturbations are neglected. This results to an error in the eigenvalues and it is to this error that bounds are derived. This is done by using the method of weighted residuals to reduce the original eigenvalue problem to that of a real asymmetric matrix. Then Gershgorin-type theorems .are used to find discs in the complex plane in which the eigenvalues are contained. The radii of the discs depend on the perturbation in a simple manner.

In the second part the effect of delayed neutrons on the eigenvalues of the group-diffusion operator is examined. The delayed neutrons cause a shifting of the prompt-neutron eigenvalue s and the appearance of the delayed eigenvalues. Using a simple perturbation method this shifting is calculated and the delayed eigenvalues are predicted with good accuracy.

Information processing and higher psychological functions in the disconnected hemispheres of human commissurotomy patients

Several patients of P. J. Vogel who had undergone cerebral commissurotomy for the control of intractable epilepsy were tested on a variety of tasks to measure aspects of cerebral organization concerned with lateralization in hemispheric function. From tests involving identification of shapes it was inferred that in the absence of the neocortical commissures, the left hemisphere still has access to certain types of information from the ipsilateral field. The major hemisphere can still make crude differentiations between various left-field stimuli, but is unable to specify exact stimulus properties. Most of the time the major hemisphere, having access to some ipsilateral stimuli, dominated the minor hemisphere in control of the body.

Competition for control of the body between the hemispheres is seen most clearly in tests of minor hemisphere language competency, in which it was determined that though the minor hemisphere does possess some minimal ability to express language, the major hemisphere prevented its expression much of the time. The right hemisphere was superior to the left in tests of perceptual visualization, and the two hemispheres appeared to use different strategies in attempting to solve the problems, namely, analysis for the left hemisphere and synthesis for the right hemisphere.

Analysis of the patients' verbal and performance I.Q.'s, as well as observations made throughout testing, suggest that the corpus callosum plays a critical role in activities that involve functions in which the minor hemisphere normally excels, that the motor expression of these functions may normally come through the major hemisphere by way of the corpus callosum.

Lateral specialization is thought to be an evolutionary adaptation which overcame problems of a functional antagonism between the abilities normally associated with the two hemispheres. The tests of perception suggested that this function lateralized into the mute hemisphere because of an active counteraction by language. This latter idea was confirmed by the finding that left-handers, in whom there is likely to be bilateral language centers, are greatly deficient on tests of perception.

Localization of sources of human evoked responses

The evoked response, a signal present in the electro-encephalogram when specific sense modalities are stimulated with brief sensory inputs, has not yet revealed as much about brain function as it apparently promised when first recorded in the late 1940's. One of the problems has been to record the responses at a large number of points on the surface of the head; thus in order to achieve greater spatial resolution than previously attained, a 50-channel recording system was designed to monitor experiments with human visually evoked responses.

Conventional voltage versus time plots of the responses were found inadequate as a means of making qualitative studies of such a large data space. This problem was solved by creating a graphical display of the responses in the form of equipotential maps of the activity at successive instants during the complete response. In order to ascertain the necessary complexity of any models of the responses, factor analytic procedures were used to show that models characterized by only five or six independent parameters could adequately represent the variability in all recording channels.

One type of equivalent source for the responses which meets these specifications is the electrostatic dipole. Two different dipole models were studied: the dipole in a homogeneous sphere and the dipole in a sphere comprised of two spherical shells (of different conductivities) concentric with and enclosing a homogeneous sphere of a third conductivity. These models were used to determine nonlinear least squares fits of dipole parameters to a given potential distribution on the surface of a spherical approximation to the head. Numerous tests of the procedures were conducted with problems having known solutions. After these theoretical studies demonstrated the applicability of the technique, the models were used to determine inverse solutions for the evoked response potentials at various times throughout the responses. It was found that reliable estimates of the location and strength of cortical activity were obtained, and that the two models differed only slightly in their inverse solutions. These techniques enabled information flow in the brain, as indicated by locations and strengths of active sites, to be followed throughout the evoked response.

Phase mapping of domain kinetics in lithium niobate by digital holographic interferometry

The phase mapping of domain kinetics under the uniform steady-state electric field is achieved and investigated in the LiNbO3 crystals by digital holographic interferometry. We obtained the sequences of reconstructed three-dimensional and two-dimensional wave-field phase distributions during the electric poling in the congruent and near stoichiometric LiNbO3 crystals. The phase mapping of individual domain nucleation and growth in the two crystals are obtained. It is found that both longitudinal and lateral domain growths are not linear during the electric poling. The phase mapping of domain wall motions in the two crystals is also obtained. Both the phase relaxation and the pinning-depinning mechanism are observed during the domain wall motion. The residual phase distribution is observed after the high-speed domain wall motion. The corresponding analyses and discussions are proposed to explain the phenomena.

Fast and Efficient Neural Conversion of Human Hematopoietic Cells

Neurons obtained directly from human somatic cells hold great promise for disease modeling and drug screening. Available protocols rely on overexpression of transcription factors using integrative vectors and are often slow, complex, and inefficient. We report a fast and efficient approach for generating induced neural cells (iNCs) directly from human hematopoietic cells using Sendai virus. Upon SOX2 and c-MYC expression, CD133-positive cord blood cells rapidly adopt a neuroepithelial morphology and exhibit high expansion capacity. Under defined neurogenic culture conditions, they express mature neuronal markers and fire spontaneous action potentials that can be modulated with neurotransmitters. SOX2 and c-MYC are also sufficient to convert peripheral blood mononuclear cells into iNCs. However, the conversion process is less efficient and resulting iNCs have limited expansion capacity and electrophysiological activity upon differentiation. Our study demonstrates rapid and efficient generation of iNCs from hematopoietic cells while underscoring the impact of target cells on conversion efficiency.

Undertreatment of human immunodeficiency virus in psychiatric inpatients: a cross-sectional study of seroprevalence and associated factors

Background: The aims of this study were to evaluate the prevalence of HIV and its associated demographic and clinical factors among psychiatric inpatients of a general hospital. Methods: This was a single-center, observational, cross-sectional study that included patients consecutively admitted to our unit aged 16 years or older and with no relevant cognitive problems. The patients were evaluated using a semistructured interview and an appropriate test for HIV infection. Results: Of the 637 patients who were screened, 546 (86%) who consented to participate were included in the analyses. Twenty-five (4.6%, 95% confidence interval [CI] 3.0-6.8) patients were HIV-positive. The prevalence was higher among patients with substance misuse (17.4%, 95% CI 9.7-28.8). All except one of the 25 patients knew of their seropositive condition prior to participation in the study. Only 14 (56%) of the 25 seropositive patients had previously received pharmacological treatment for their infection. According to the multiple logistic regression analysis, the likelihood of HIV infection was lower in patients with higher levels of education and higher among patients who were single, had history of intravenous drug use, and had an HIV-positive partner, particularly if they did not use condoms. Among the patients with HIV infection, 18 (72%) had a history of suicide attempts compared with 181 (34.7%) of the patients without HIV infection (relative risk 2.1, 95% CI 1.6-2.7; P<0.001). Conclusion: HIV infection is highly prevalent in patients admitted to a psychiatric unit, especially those with a diagnosis of substance misuse. Seropositive patients show very poor treatment adherence. The risk of suicide seems to be very high in this population. Implementing interventions to reduce the suicide risk and improve adherence to antiretroviral therapy and psychotropic medications seems crucial.